Made available in DSpace on 2015-01-26T13:21:29Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-08-15Bitstream added on 2015-01-26T13:30:22Z : No. of bitstreams: 1 000792740.pdf: 3309974 bytes, checksum: 896459872b09865fb008a59c2ddbe95c (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / A família Sapindaceae é bastante estudada do ponto de vista farmacológico. A Serjania marginata Casar., uma espécie dessa família, é indicada, popularmente, para dores de estômago. Os objetivos deste trabalho foram estabelecer o perfil químico, isolar e caracterizar os constituintes químicos do extrato EtOH 70% de S. marginata, quantificar as substâncias marcadoras da espécie, avaliar a toxicidade aguda, neurotoxicidade, atividade anti-ulcerogênica, antiradicalar e mutagênica do extrato, determinar o teor de fenóis e flavonoides totais e avaliar a espécie no cultivo ex situ. Os estudos foram realizados com as folhas de plantas nativas do Cerrado e cultivadas no Horto de Plantas Medicinais, da UFGD, em Dourados-MS. O maior rendimento foi da espécie nativa coletada em fevereiro de 2011 (32,6%). Pela triagem fitoquímica, determinou-se que o extrato é composto por flavonoides, saponinas, taninos, compostos fenólicos e esteróides. Foram isoladas quinze substâncias, das classes dos flavonoides, taninos e saponinas, sendo o ácido oleanólico 3-O-α-L-arabinopiranosil(1→3)-α-L-ramnopiranosil(1→2)[β-D-glicopiranosil(1→4)]-α-L-arabinopiranosídeo e a 7,5”-anidroapigenina 8-C-α-(2,6-dideoxi-5-hidroxi-ribo-hexopiranosil)-4'-O-β-D-glucopiranosídeo, descritos pela primeira vez na literatura. O extrato não apresentou efeitos tóxicos nem neurotóxicos no uso oral agudo e não foi mutagênico. Evidenciou-se também significativa atividade anti-úlcera para o extrato em diversos modelos farmacológicos, avaliados in vivo. Os extratos das folhas colhidas em diferentes épocas de cultivo, avaliados através do uso do reagente DPPH, apresentaram alta atividade antiradicalar, quando comparados ao ácido gálico e quercetina, especialmente devido à presença de taninos. As plantas coletadas após o florescimento apresentaram maior quantidade de compostos fenólicos e, antes do florescimento, maior concentração de flavonoides... / Sapindaceae family is extensively studied on a pharmacological point of view. Serjania marginata Casar., a specie of this family, is indicated popularly for stomach aches. The objectives of this work were to standardize the chemical profile, to characterize the chemical composition of 70% EtOH extract from S. marginata, to quantify the marker substances for the specie, to assess the acute toxicity, neurotoxicity, anti-ulcerogenic, antiradical and mutagenic activities of the extract, to determine the total phenolic and flavonoid contents and to evaluate conditions of ex situ cultivation for the specie. The studies were performed with leaves of native plants from the Cerrado and of plants grown in the Garden of Medicinal Plants from the UFGD, Dourados-MS. The highest contents were achieved by the native accesses collected during February 2011 (32.6%). During the chemical screening, it was determined that the extract was composed by flavonoids, saponins, tannins, phenols and steroids. Fifteen substances were isolated which were classified as flavonoids, tannins and saponins, and the oleanolic acid 3-O-α-L-arabinopyranosyl(1→3)-α-L-rhamnopyranosyl(1→2)[β-D-glucopyranosyl(1→4)]-α-L-arabinopyranoside and 7,5”-anhidroapigenin 8-C-α-(2,6-dideoxy-5-hydroxy-ribo-hexopyranosyl)-4'-O-β-D-glucopyranoside described for the first time in the literature. The extract here studied did not show any toxic or neurotoxic effect by acute oral dose neither presented any mutagenic potential. Significant anti-ulcer activity of the extract was evidenced when applied in different pharmacological modulations, evaluated in vivo. The extracts from leaves harvested in different growing seasons evaluated through the use of the reagent DPPH showed high antiradical activity when compared to the gallic acid and quercetin, mainly due to the presence of tannins. Plants collected after flowering showed higher amounts of phenolic compounds and, before flowering, ... / FAPESP: 11/08880-4

Overweight and obesity are becoming a major public health issue. Pancreatic lipase is a key enzyme to catalyze the hydrolysis of 50-70% of dietary fat in the digestive system. Inhibition of pancreatic lipase activity can block fat absorption in gastrointestinal tract, further control obesity incidence. On the other hand, a-glucosidase is also a key enzyme to hydrolysis polysaccharides and disaccharides into glucose in small intestine. Inhibition of a-glucosidase activity can block digestion and absorption of carbohydrates, further control metabolism disorders such as diabetes and obesity. Food legumes are widely used in people’s life with multiple pharmacological activities. The author extracted total phenolics and saponin components from 13 commonly consumed food legumes produced in China, did a systematic comparative study investigating their inhibitory effects against digestive enzymes (pancreatic lipase and a-glucosidase), and screened adzuki bean (Vigna angularis L.) as the further target bean. The results showed that the different concentrations of total phenolic extract from adzuki bean (0.25, 0.5, 0.75, 1 mg/mL) inhibited hydrolysis of triolein about 24.1, 24.0, 30.7, and 36.3%, respectively, while different concentrations of total saponin extracts from adzuki bean (0.25, 0.5, 0.75, 1 mg/mL) inhibited hydrolysis of triolein about 15.9, 23.5, 30.1, and 29.2%, respectively. On the other hand, phenolic extract of adzuki bean at the concentration from 0.25 to 1 mg/mL exhibited much more than 80% a-glucosidase inhibitory activity, while saponin extract of adzuki bean exhibited 56.4 to 68.7% a-glucosidase inhibitory activity. Based on the results of pancreatic lipase inhibitory activity, at the concentration of 1 mg/mL, phenolics extract and saponin extract of adzuki bean (mean 32.5%) and pinto bean (mean 27%) had the stronger inhibitory effects. Moreover, phenolics extract and saponin extract of adzuki bean (mean 79.25%) and pinto bean (mean 72.85%) also had the stronger inhibitory effects on a-glucosidase activity. In addition, adzuki bean is widely used and has diverse application in foods and drugs. In a word, it indicated that adzuki bean is one of the best target beans to further study anti-obesity and anti-diabetes effects via cell and animal models. Flavonoids and saponins in adzuki bean were obtained and characterized by high performance liquid chromatography with diode array detection and electro spray ionization-tandem multi-stage mass spectrometry in succession. Among 15 compounds identified, four flavonoids (catechin, vitexin-4.-O-glucoside, quercetin-3-O-glucoside, and quercetin-3-O-rutinoside) and six saponins (azukisaponin I, II, III, IV, V, and VI) in adzuki bean were further quantified by external calibration method using high performance liquid chromatography mass spectrometry with the program of time segment and extract ion chromatogram analysis. Animal model is a good way to intuitively evaluate the anti-obesity effect of adzuki bean. So the anti-obesity effects of adzuki bean in mice fed with a high-fat diet was investigated. ICR female mice were fed with a high fat diet administrated orally with different doses of adzuki bean extracts for eight weeks. Total extract, flavonoids and saponins of adzuki bean enhanced lipolysis (166.1%, 175.6%, and 152.6%, respectively). Compared to the final body weight (33.6 g) of the high-fat diet group, oral administration (300 mg/kg per day) of total extract, flavonoids and saponins of adzuki bean significantly reduced the final body weight of mice, and significantly decreased the adipose tissue accumulation. The adzuki bean intervention also significantly reduced the levels of serum triglyceride, total cholesterol, low density lipoprotein-cholesterol, and liver lipid. Adzuki bean demonstrated the anti-obesity effects on mice fed with a high fat diet, such effects may mediated through the inhibitory effects of flavonoids and saponins from adzuki bean on a-glucosidase and pancreatic lipase activities, and lipolysis enhancement effect of active components from adzuki bean. Obesity is characterized biologically at the cellular level by as an increase in the number of preadipocutes and an increase in the size of adipocytes differentiated from preadipocytes in adipose tissue. 3T3-L1 cell model was used to reveal the mechanism of anti-obesity effects of adzuki bean in the present study. The results showed that adzuki bean total extract, adzuki bean flavonoids, adzuki bean saponins, four mono flavonoides (catechin, vitexin-4.-O-glucoside, quercetin-3-O-glucoside, and quercetin-3-O-rutinoside) and six mono saponins (azukisaponin I, II, III, IV, V, and VI) exhibited inhibitory effect of proliferation of 3T3-L1 cells, and the inhibitory rate of proliferation of 3T3-L1 cells was about 20%. Mature adipocytes were stained by Oil Red O and the lipid accumulated exhibited red color, while the preadipocytes could not been stained. The data of the effects of adzuki bean samples on lipid accumulation during the differentiation period of 3T3-L1 cells revealed that azukisaponin II had the highest inhibitory effect (49.72%) with a dose dependent manner from 50 to 500 µg/mL. It indicated that adzuki bean may block the differentiation of 3T3-L1 cells from preadipocytes to mature adipocytes which contribute to anti-obesity effects. Most of adzuki bean samples significantly blocked the triglyceride accumulation and enhanced lipolysis by increseaing the released of glycerol during differentiation of 3T3-L1 cell. Additionally, adzuki bean samples except catechin significantly inhibited glycerol-3-phosphase dehydrogenase activity in 3T3-L1 cells, which indicated that adzuki bean samples had the ability to inhibit triglyceride synthesis. Real-time polymerase chain reaction and Western blot methods were used to investigate the intervention effects of adzuki bean total extract, adzuki bean flavonoids, adzuki bean saponins, quercetin-3-O-rutinoside, and azukisaponin II isolated from adzuki bean on the related gene expressions. These adzuki bean samples inhibited differentiation of 3T3-L1 cell by decreasing the expression of peroxisome proliferator-activated receptor . and CCATT/enhancer-binding protein a (major adipocyte transcription factors) at messenger ribonucleic acid level and protein level in adipocytes. Additional, they inhibited adipogenesis by decreasing the expression of fatty acid synthase, promoting lipolysis by increasing the expression of hormone-sensitive lipase and adipose triglyceride lipase, made energy balance by regulating the expression of leptin, adenosine monophosphate -activated protein kinase, and tumor necrosis factor a, and induced cell apoptosis by increasing the expression of bax and decresing the expression of B-cell lymphoma 2. These findings provide insight into the molecular mechanisums through regulation of the related gene expression in 3T3-L1 adipocytes. It also suggested that adzuki bean had a positive effect in prevention and treatment of adipogenesis-related obesity, and might be a good option of functional foods to control obesity.

Made available in DSpace on 2014-06-11T19:27:28Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-02-12Bitstream added on 2014-06-13T20:56:35Z : No. of bitstreams: 1 oliveira_aps_me_arafcf.pdf: 946584 bytes, checksum: 6c4fbb82549dc1046e8b7771847a8f5b (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / A família Eriocaulaceae possui cerca de 1200 espécies divididas em 10 gêneros e suas espécies de plantas são conhecidas como sempre-vivas, por sua grande durabilidade e coloração paleácea. Neste trabalho foram estudadas as espécies: Eriocaulon ligulatum, Leiothrix flavescens e Leiothrix spiralis, as quais foram avaliadas quanto à mutagenicidade e estrogenicidade e algumas substâncias isoladas foram utilizadas para um estudo de relação estrutura-atividade mutagênica e estrogênica. Para avaliação da mutagenicidade, utilizou-se o teste de Ames, e o teste RYA (Recombinant Yeast Assay) foi o empregado para avaliação da estrogenicidade. Os resultados da atividade mutagênica demonstraram que apenas E. ligulatum foi considerado mutagênico e isso foi atribuído às isocumarinas e flavonóides agliconas. Os estudos de relação estrutura-atividade mutagênica mostraram que esta ação foi devida à presença de hidroxilas em posições estratégicas e à ausência de glicosilações, metoxilações, ou qualquer outro tipo de substituição. Para complementar os estudos de relação estrutura-atividade mutagênica com as isocumarinas, utilizou-se resultados já publicados de mutagenicidade de moléculas semelhantes à isocumarina eriocaulina, isolada de E. ligulatum. Verificou-se que os dímeros de isocumarinas têm a sua mutagenicidade influenciada pela conformação espacial, pela presença de grupos hidroxilas mais afastados de grupos volumosos, e pelo tipo de ligação que une os seus monômeros. Quando o potencial estrogênico foi avaliado, apenas a L. flavescens apresentou resultados positivos e que esses são devidos à presença de flavonas agliconas, como a luteolina e a metoxiluteolina. As mesmas isocumarinas, pertencentes a outras espécies da família Eriocaulaceae, avaliadas no teste de Ames, foram avaliadas no teste RYA... / Eriocaulaceae family comprises around 1200 species, divided into 10 genera, and their plant species are known as “sempre-vivas” (everlasting flowers) for its durability and color. In this work the species Eriocaulon ligulatum, Leiothrix flavescens and Leiothrix spiralis were studied. These species were evaluated for the mutagenic and the estrogenic potential and some of their isolated compounds were used for a study of structuremutagenic and estrogenic activities. To assess mutagenicity we used Ames test and the Recombinant Yeast Assay (RYA test) was used for evaluation of estrogenicity. The results of mutagenic activity showed that only E. ligulatum presented mutagenicity and that these results were due to its naphthopyranones and flavonoid aglicons. The structuremutagenic activity studies shown that this action were due to the presence of hydroxyl groups in strategic positions and no glycosylations, methoxylation or any type of substitutions. To complement the studies of structure-mutagenic activity with the naphthopyranones, we use results already published of the mutagenicity of similar compounds to the naphthopyranone eriocaulina, isolated from E. ligulatum, and compare them with the data from ericauline. It was found that the dimers of naphthopyranones have their mutagenicity influenced by its spatial conformation, for the presence of hydroxyl groups furthest away from large groups, and the type of connection that join their monomers. When the estrogenic potential was assessed, we found that only L. flavescens showed positive results and that these results are due to the presence of flavones in the form of aglycones, such as luteolin and methoxyluteolin. The same naphthopyranones belonging to other species of Eriocaulaceae family, evaluated in the Ames test, were assessed in the RYA test. Their results were crucial for the establishment of structureestrogenic activity... (Complete abstract click electronic access below)

Submitted by LUIS FERNANDO MERCES CHAVES SILVA null (luisfernando.mchaves@gmail.com) on 2016-06-02T00:52:25Z No. of bitstreams: 1 TESE MESTRADO VERSÃO FINAL (LUIS FERNANDO M. C. SILVA).pdf: 2997186 bytes, checksum: 711036d97c3d4d1e8cce675ab6ad1477 (MD5) / Approved for entry into archive by Juliano Benedito Ferreira (julianoferreira@reitoria.unesp.br) on 2016-06-06T14:47:13Z (GMT) No. of bitstreams: 1 silva_lfmc_me_bot.pdf: 2997186 bytes, checksum: 711036d97c3d4d1e8cce675ab6ad1477 (MD5) / Made available in DSpace on 2016-06-06T14:47:13Z (GMT). No. of bitstreams: 1 silva_lfmc_me_bot.pdf: 2997186 bytes, checksum: 711036d97c3d4d1e8cce675ab6ad1477 (MD5) Previous issue date: 2016-04-27 / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / O surgimento das biotecnologias aplicadas ao sêmen equino, como a refrigeração e congelação, representaram um grande avanço para equinocultura, principalmente por permitir um melhor aproveitamento do ejaculado, otimizando a utilização de animais considerados geneticamente superiores. Entretanto a aplicação dessas técnicas imprime estresse térmico às células espermáticas, resultando em injúrias físicas e químicas, dentre elas uma elevação significativa na produção de espécies reativas do oxigênio (EROs). Quando em excesso, estes agentes instáveis são capazes de reagir e promover lesão às enzimas, lipídeos de membrana e ácidos nucleicos, reduzindo a viabilidade dos espermatozoides criopreservados. Desta maneira surge a necessidade de se introduzir elementos que desempenhem função antioxidante nos meios de refrigeração e congelação. A quercertina, um polifenol do grupo dos flavonoides, é considerada um poderoso antioxidante por reduzir o surgimento das principais EROs, quelar íons metálicos formadores de radicais livres e estabilizar a peroxidação lipídica. Apesar disto, a eficiência da aplicação deste antioxidante sobre o ejaculado equino durante o processo de criopreservação ainda é controversa e pouco referenciada, sendo necessário mais estudos para que se determine a influência de sua utilização no ejaculado de garanhões durante a refrigeração e congelação. / The emergence of biotechnologies applied to the equine semen as cooling and freezing process represented a major breakthrough for Equine, especially for allowing a better use of the ejaculate, optimizing the use of genetically superior animals. However the application of these techniques induces thermal stress on sperm cells, resulting in physical and chemical injuries, among them a significant rise in the production of reactive oxygen species (ROS). These unstable agents when in excess, are able to react and promote damage to enzymes, lipid membrane and nucleic acids, thereby reducing the viability of cryopreserved sperm. Thus is required to introduce elements that perform antioxidant function in cooling and freezing extenders. The quercertina, a polyphenol of the flavonoids group, is considered a potent antioxidant by reducing the emergence of the main ROS, chelate metal ions producers of free radicals and stabilize the lipid peroxidation. Despite this, the efficiency of application of this antioxidant on the equine ejaculate during the process of cryopreservation is still controversial and poorly referenced, requiring further studies in order to determine if there is influence of their use in the ejaculate of stallions during cooling and freezing. / FAPESP: 2014/01681-4

Made available in DSpace on 2014-06-11T19:29:10Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-02-19Bitstream added on 2014-06-13T20:38:29Z : No. of bitstreams: 1 moura_acs_me_araiq.pdf: 1236232 bytes, checksum: a709145cb5de5a7efb7d91cdfaf1362f (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / As plantas medicinais utilizadas no tratamento de diversas doenças são fontes de uma infinidade de novas moléculas de interesse para a humanidade. Este trabalho investigou a composição química das partes aéreas do chá e do extrato metanólico de Indigofera hirsuta (Fabaceae), popularmente conhecida como “anileira” e encontrada no Cerrado brasileiro. O chá foi fracionado por cromatografia de permeação em gel e cromatografia líquida de média pressão, tendo-se isolado: kaempferol, ácido protocatecuico, kaempferol-3-O-β-D-glucopiranosideo, triptofano e uracila. O extrato metanólico foi fracionado por cromatografia líquida de alta pressão, que possibilitou o isolamento de: ácido gálico, galato de metila, rutina e kaempferol-3-O-β-D-glucopiranosideo. As substâncias foram identificadas por técnicas espectroscópicas (UV, IV e RMN) e as estruturas confirmadas pelo uso da técnica acoplada de HPLC-DAD-ESI-MS. O chá, extrato metanólico e as substâncias isoladas foram avaliados quanto à atividade antioxidante, utilizando-se DPPH. O extrato metanólico apresentou significativa atividade antioxidante, enquanto que o chá não apresentou atividade antioxidante considerável, quando comparados com o padrão ácido gálico. Das substâncias isoladas, o galato de metila apresentou a maior atividade antioxidante. O chá de I. hirsuta foi também avaliado quanto à atividade gastroprotetora em lesões induzida por etanol absoluto em 3 diferentes doses: 125 mg.Kg-1, 250 mg.Kg-1 e 500 mg.Kg-1, sendo a menor dose a que apresentou melhor efeito. Contudo, o chá de Indigofera hirsuta não apresentou atividade gastroprotetora / Medicinal plants used in the treatment of various diseases and are source of many new molecules. This work investigates the chemical composition of the aerial parts of the infusion and methanol extract of Indigofera hirsuta (Fabaceae), popularly known as “anileira” and found in the Brazilian Cerrado. The infusion was fractionated by gel permeation chromatography and medium pressure liquid chromatography, it was possible to isolate kaempferol, protocatecuic acid, kaempferol-3-O-β-D-glucopyranoside, tryptophan and uracil. The methanol extract was fractionated by high pressure liquid chromatography, which allowed the isolation of gallic acid, methyl gallate, rutin and kaempferol-3-O-β-D-glucopyranoside. The substances were identified by spectroscopic techniques (UV, IR and NMR) and confirmed by the use of HPLC-DAD-ESI-MS coupling. The infusion, methanol extract and isolated compounds were evaluated for their antioxidant activity using DPPH. The methanol extract showed significant antioxidant activity, while the tea did not show significant antioxidant activity, when compared to gallic acid. Among the isolated substances methyl gallate presented the highest antioxidant activity. The I. hirsuta was also evaluated for its gastroprotective activity in lesions induced by absolute ethanol in 3 different doses (125 mg.Kg-1, 250 mg.Kg-1 and 500 mg.Kg-1).The lowest dose showed the best effect. However, the infusion not active gastroprotective

Resumo: A etiologia da colite ulcerativa ainda permanece desconhecida, porém atinge uma grande parcela da população mundial. A maioria das terapias disponiveis tem eficácia limitada e geram significativos clínicos efeitos adversos. A especie medicinal Mouriri pusa Gardn. (Melastomataceae) foi citada diversas vezes pela população local do Tocantins como útil no tratamento de distúrbios do trato digestório. Os estudos preliminares com essa planta tem apresentado efeitos gastroprotetores e cicatrizantes intensos. Estudos fitoquimicos de seu extrato metanólico revelaram presença de (-)- epicatequina, além de outros flavonóides, e taninos. Nesse projeto, induziu-se colite ulcerativa em ratos per injeção local de acido trinitrobenzenosulfonico (TNBS). Foram analisados parametros morfológicos, inflamatórios, imunohistoquimicos e Western Blot. 0 efeito da (-)-epicatequina foi avaliado em modele agudo nas doses de 5,10,25 e 50 mg/kg (p.o). Os animais receberam os tratamentos uma vez por dia durante 3 dias antes da indução de colite, 2 h e 24 h após a mesma. Observou-se macroscopicamente que as doses de 10 e 50 mg/kg (EC10 e ECSO)de epicatequina foram efetivas em reduzir 0 grau de seriedade da lesão, sendo confirmado pela análise microscópica para EC10. Nas quantificações do tecido intestinal, observou-se que a concentração de glutationa em EC10 foi maior que no controle, mostrando um possivel mecanismo de ação, já que a manutenção da glutationa no tecido inflamado pode indicar potencial antioxidante da substância. Observou-se diminuição de COX-2, um parametro inflamatório, e um aumento de PCNA, antigeno de proliferação celular, em EC10, dicando ação antiinflamatória e estimuladora de proliferação celular. EC10 e EC50 foram usadas então para o modelo crônico de colite, no qual os diferentes grupos de animais receberam, após indução de colite no primeiro dia ... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Ulcerative colitis etiology is still not completely known, however, it is present among great portion of world's population. The majority of available therapies has limited efficacy and provides significant adverse effects. The medicinal species Mouriri pusa Gardn. (Melastomataceae) has been often mentioned by local population from Tocantins as useful in the treatment of gastrointestinal disorders. Previous studies with this plant have presented intense gastroprotective and cicatrizing effects. Phyochemical studies of its methanolic extract revealed presence of (-)-epicatechin, besides other flavonoids, and tannins, In this work, ulcerative colitis was induced in rats by local injection of TNBS (Trinitrobenzenesulphonic acid). We analyzed morphological, inflammatory, immunohistochemical and Western Blot parameters. (-)- Epicatechin effect was evaluated in acute model in 5, 10, 25 e 5Omg/kg doses (p.o). The animals received treatments once a day for 3 days before colitis induction, and 2 h and 24 h after it. The doses of 10 and 5Omg/kg (EC10 and EC5O) were macroscopically observed to be effective in reducing lesion seriousness, what was confirmed by microscopical analysis for EC10. By quantifications in the colon tissue, it was observed that glutathion concentration in EC10 was greater than in control, showing a possible echanism of action, since glutathione maintenance in the inflamed tissue may . dicate antioxidant potential from the compound. It was observed COX-2 reduction, an inflammatory parameter, and an increase in PCNA, proliferating cell antigen, in EClO, indicating anti-inflammatory and proliferation stimulating action. EC10 and EC were then used for chronic colitis model, in which the groups, after colitis induction in the first Day, treatments for 21 days. At the end of each week, some of the rats from each group were killed for accompaniment, and after the second week ... (Complete abstract click electronic access below) / Orientador: Cláudia Helena Pellizzon / Coorientador: Clélia Akiko Hiruma-Lima / Banca: Luís Cláudio Di Stasi / Banca: João Carlos Palazzo de Mello / Mestre

Orientador : Wagner Vilegas / Banca: Ana María Simonet Morales / Banca: Cássia Regina Primila Cardoso / Banca: Luís Vitor Silva do Sacramento / Banca: Marcos Hikari Toyama / Resumo: A família Sapindaceae é bastante estudada do ponto de vista farmacológico. A Serjania marginata Casar., uma espécie dessa família, é indicada, popularmente, para dores de estômago. Os objetivos deste trabalho foram estabelecer o perfil químico, isolar e caracterizar os constituintes químicos do extrato EtOH 70% de S. marginata, quantificar as substâncias marcadoras da espécie, avaliar a toxicidade aguda, neurotoxicidade, atividade anti-ulcerogênica, antiradicalar e mutagênica do extrato, determinar o teor de fenóis e flavonoides totais e avaliar a espécie no cultivo ex situ. Os estudos foram realizados com as folhas de plantas nativas do Cerrado e cultivadas no Horto de Plantas Medicinais, da UFGD, em Dourados-MS. O maior rendimento foi da espécie nativa coletada em fevereiro de 2011 (32,6%). Pela triagem fitoquímica, determinou-se que o extrato é composto por flavonoides, saponinas, taninos, compostos fenólicos e esteróides. Foram isoladas quinze substâncias, das classes dos flavonoides, taninos e saponinas, sendo o ácido oleanólico 3-O-α-L-arabinopiranosil(1→3)-α-L-ramnopiranosil(1→2)[β-D-glicopiranosil(1→4)]-α-L-arabinopiranosídeo e a 7,5"-anidroapigenina 8-C-α-(2,6-dideoxi-5-hidroxi-ribo-hexopiranosil)-4'-O-β-D-glucopiranosídeo, descritos pela primeira vez na literatura. O extrato não apresentou efeitos tóxicos nem neurotóxicos no uso oral agudo e não foi mutagênico. Evidenciou-se também significativa atividade anti-úlcera para o extrato em diversos modelos farmacológicos, avaliados in vivo. Os extratos das folhas colhidas em diferentes épocas de cultivo, avaliados através do uso do reagente DPPH, apresentaram alta atividade antiradicalar, quando comparados ao ácido gálico e quercetina, especialmente devido à presença de taninos. As plantas coletadas após o florescimento apresentaram maior quantidade de compostos fenólicos e, antes do florescimento, maior concentração de flavonoides... / Abstract: Sapindaceae family is extensively studied on a pharmacological point of view. Serjania marginata Casar., a specie of this family, is indicated popularly for stomach aches. The objectives of this work were to standardize the chemical profile, to characterize the chemical composition of 70% EtOH extract from S. marginata, to quantify the marker substances for the specie, to assess the acute toxicity, neurotoxicity, anti-ulcerogenic, antiradical and mutagenic activities of the extract, to determine the total phenolic and flavonoid contents and to evaluate conditions of ex situ cultivation for the specie. The studies were performed with leaves of native plants from the Cerrado and of plants grown in the Garden of Medicinal Plants from the UFGD, Dourados-MS. The highest contents were achieved by the native accesses collected during February 2011 (32.6%). During the chemical screening, it was determined that the extract was composed by flavonoids, saponins, tannins, phenols and steroids. Fifteen substances were isolated which were classified as flavonoids, tannins and saponins, and the oleanolic acid 3-O-α-L-arabinopyranosyl(1→3)-α-L-rhamnopyranosyl(1→2)[β-D-glucopyranosyl(1→4)]-α-L-arabinopyranoside and 7,5"-anhidroapigenin 8-C-α-(2,6-dideoxy-5-hydroxy-ribo-hexopyranosyl)-4'-O-β-D-glucopyranoside described for the first time in the literature. The extract here studied did not show any toxic or neurotoxic effect by acute oral dose neither presented any mutagenic potential. Significant anti-ulcer activity of the extract was evidenced when applied in different pharmacological modulations, evaluated in vivo. The extracts from leaves harvested in different growing seasons evaluated through the use of the reagent DPPH showed high antiradical activity when compared to the gallic acid and quercetin, mainly due to the presence of tannins. Plants collected after flowering showed higher amounts of phenolic compounds and, before flowering, ... / Doutor

Orientador: Wagner Vilegas / Coorientador: Miriam Sannomiya / Banca: Alba Regina Monteiro Souza Brito / Banca: Elfriede Marianne Bacchi / Resumo: O uso de fitoterápicos padronizados com assegurada comprovação de eficácia e segurança tem apresentado resultados terapêuticos surpreendentes em diversos países. Contudo, no Brasil, onde há grande potencial de produção de fitoterápicos devido a sua enorme biodiversidade, há deficiências na correta caracterização química das matérias-primas e escassez de ensaios farmacológicos e toxicológicos desses materiais, etapas essenciais para assegurar eficácia e segurança dos fitoterápicos. Machaerium hirtum (Fabaceae) é utilizada popularmente para o tratamento de diarréia, tosse e câncer. Um estudo químico que identifique as substâncias presentes nesta espécie e as quantifique pode fornecer subsídios para o entendimento destas atividades. A primeira etapa consistiu no isolamento das substâncias do extrato EtOH 70% das partes aéreas de M. hirtum por meio de cromatografia de permeação em gel seguida de purificação das substâncias por HPLC-RI. A identificação das substâncias foi feita por Ressonância Magnética Nuclear e por Espectrometria de Massas. O perfil químico do extrato foi estabelecido por HPLC-PDA e a quantificação das substâncias foi feita com base em suas agliconas, pelo método da calibração externa. Foi possível identificar as flavonas saponarina (8,69 ± 0,46 mg/g de extrato), isovitexina (6,44 ± 0,45 mg/g de extrato), isoorientina, que se apresentou abaixo do limite de quantificação (LOQ) e swertisina, presente no perfil cromatográfico em co-eluição com outra substância. Tais resultados são fundamentais para a padronização do extrato EtOH 70% das folhas de Machaerium hirtum / Abstract: The use of standardized phytomedicines with proven efficacy and safety has shown surprising therapeutic results in various countries. Brazil has great potential for production of herbal medicine due to its enormous biodiversity. However, there are deficiencies in the correct characterization of chemical raw materials and few reliable pharmacological and toxicological tests of these materials, essential steps to ensure efficacy and safety of herbal medicines. Machaerium hirtum (Fabaceae) is popularly used to treat diarrhea, cough and cancer. A chemical study to identify the substances present in this species may provide and quantify contributions to the understanding of these activities. The first step was the isolation of substances of 70% EtOH extract of aerial parts of M. hirtum by gel permeation chromatography followed by purification of substances by HPLC-RI. The identification of substances was performed by Nuclear Magnetic Resonance and Mass Spectrometry. The chemical profile of the extract was established by HPLC-PDA and quantification of the substances was based on their aglycones by the method of external calibration. It was possible to identify the flavones saponarin (8,69 ± 0,46 mg/g of extract), isovitexin (6,44 ± 0,45 mg/g of extract), isoorientin, which was below the limit of quantification (LOQ) and swertisin, present in the chromatographic profile in co-elution with another substance. These results are fundamental to the standardization of the crude 70% EtOH extract from the leaves of Machaerium hirtum / Mestre

Orientador: Frederico Ozanam Papa / Resumo: O surgimento das biotecnologias aplicadas ao sêmen equino, como a refrigeração e congelação, representaram um grande avanço para equinocultura, principalmente por permitir um melhor aproveitamento do ejaculado, otimizando a utilização de animais considerados geneticamente superiores. Entretanto a aplicação dessas técnicas imprime estresse térmico às células espermáticas, resultando em injúrias físicas e químicas, dentre elas uma elevação significativa na produção de espécies reativas do oxigênio (EROs). Quando em excesso, estes agentes instáveis são capazes de reagir e promover lesão às enzimas, lipídeos de membrana e ácidos nucleicos, reduzindo a viabilidade dos espermatozoides criopreservados. Desta maneira surge a necessidade de se introduzir elementos que desempenhem função antioxidante nos meios de refrigeração e congelação. A quercertina, um polifenol do grupo dos flavonoides, é considerada um poderoso antioxidante por reduzir o surgimento das principais EROs, quelar íons metálicos formadores de radicais livres e estabilizar a peroxidação lipídica. Apesar disto, a eficiência da aplicação deste antioxidante sobre o ejaculado equino durante o processo de criopreservação ainda é controversa e pouco referenciada, sendo necessário mais estudos para que se determine a influência de sua utilização no ejaculado de garanhões durante a refrigeração e congelação. / Abstract: The emergence of biotechnologies applied to the equine semen as cooling and freezing process represented a major breakthrough for Equine, especially for allowing a better use of the ejaculate, optimizing the use of genetically superior animals. However the application of these techniques induces thermal stress on sperm cells, resulting in physical and chemical injuries, among them a significant rise in the production of reactive oxygen species (ROS). These unstable agents when in excess, are able to react and promote damage to enzymes, lipid membrane and nucleic acids, thereby reducing the viability of cryopreserved sperm. Thus is required to introduce elements that perform antioxidant function in cooling and freezing extenders. The quercertina, a polyphenol of the flavonoids group, is considered a potent antioxidant by reducing the emergence of the main ROS, chelate metal ions producers of free radicals and stabilize the lipid peroxidation. Despite this, the efficiency of application of this antioxidant on the equine ejaculate during the process of cryopreservation is still controversial and poorly referenced, requiring further studies in order to determine if there is influence of their use in the ejaculate of stallions during cooling and freezing. / Mestre

Embora seja uma das áreas mais ameaçadas na Terra, há poucos estudos sobre o potencial farmacológico da Caatinga. Este bioma é exclusivamente brasileiro e as plantas encontradas na região apresentam características únicas que as tornam fontes imensuráveis de compostos ativos. O parasito Trichomonas vaginalis é o agente etiológico da tricomoníase, a doença sexualmente transmissível não viral mais comum no mundo. A infecção está associada a sérias consequências em saúde e casos de resistência ao fármaco de escolha, metronidazol, estão em constante crescimento. Desta forma, a necessidade por alternativas para o tratamento da tricomoníase é evidente. O objetivo desta tese foi avaliar o potencial anti-T. vaginalis de plantas oriundas da Caatinga. A planta Manilkara rufula apresentou a mais potente atividade frente ao parasito e um fracionamento bioguiado foi realizado. Triterpenos pentacíclicos (caproato de α- e β-amirina, acetato de β-amirina e lupeol) foram identificados nas frações apolares de M. rufula, porém com baixa atividade antiparasitária. Derivados triterpênicos semissintéticos, frações contendo flavonoides, taninos e saponinas também foram testados. Nossos resultados demonstraram que o ácido ursólico e a fração H100 apresentaram potente atividade anti-T. vaginalis e ambos afetaram o crescimento e viabilidade dos trofozoítos. Além disso, o ácido ursólico e a fração H100 induziram importantes alterações na ultraestrutura dos organismos, indicando que o mecanismo de morte do parasito ocorreu por danos na membrana. A fração H100 induziu a lise total de eritrócitos e reduziu a adesão dos parasitos às células epiteliais, sugerindo, novamente, a ação sobre membrana. A fração H100 não apresentou citotoxicidade às células epiteliais. Os resultados apresentados demonstraram o potencial da planta M. rufula contra T. vaginalis e contribuem para o entendimento das propriedades farmacológicas das plantas da Caatinga, bem como para o desenvolvimento racional de alternativas para o tratamento da tricomoníase. / Although it is one of the most endangered areas on Earth, there are few studies on the pharmacological potential of the Caatinga. This is an exclusively Brazilian biome and its plants present unique characteristic that make them an immeasurable source of active compounds. The parasite Trichomonas vaginalis is the etiologic agent of trichomoniasis, the most common non-viral sexually transmitted disease worldwide. The infection has been associated with serious consequences in health and cases of resistance to metronidazole are constantly increasing. Then, the necessity of new alternatives to the treatment of trichomoniasis is evident. The aim of this thesis was to evaluate the anti-T. vaginalis potential of plants from Caatinga. The Manilkara rufula plant showed the most potent activity against the parasite and bioguided fractionation was performed. Pentacyclic triterpenes (α- and β-amyrin caproate and β-amyrin and lupeol acetate) were identified in the nonpolar fractions of M. rufula, however, low antiparasitic activity was observed. Triterpenic semisynthetic derivatives, fractions containing flavonoids, tannins and saponins were also tested. Our results showed that ursolic acid and H100 fraction exhibited potent anti-T. vaginalis activity and both affect growth and viability of trophozoites. In addition, ursolic acid and H100 fraction induced significant alterations in the ultrastructure of the parasite, indicating that the mechanism of death is by membrane damage. The H100 fraction induced complete lysis of erythrocytes and reduced the parasites adhesion to epithelial cells, once again, suggesting that the H100 action was on parasite membrane. The fraction H100 showed no cytotoxicity to epithelial cells. The results demonstrated the potential of the plant M. rufula against T. vaginalis and contributed to the understanding of the pharmacological properties of the Caatinga plants, as well as for the rational development of alternatives for trichomoniasis treatment.